猫传染性腹膜炎病毒荧光定量RT-PCR检测方法的建立  

Establishment of a Quantitative RT-PCR Assay for Feline Infectious Peritonitis Virus

作　　者：杨妮 韩佃刚 杨云庆 叶玲玲 李静 周思佳 宿放 艾军 信吉阁 Yang Ni;Han Diangang;Yang Yunqing;Ye Lingling;Li Jing;Zhou Sijia;Su Fang;Ai Jun;Xin Jige(College of Veterinary Medicine,Yunnan Agricultural University,Kunming,Yunnan 650201,China;Technology Center of Kunming Customs,Kunming,Yunnan 650200,China)

机构地区：[1]云南农业大学动物医学院,云南昆明650201 [2]昆明海关技术中心,云南昆明650200

出　　处：《中国动物检疫》2022年第10期127-131,共5页China Animal Health Inspection

基　　金：云南省院士专家工作站项目(2018IC078);兴滇英才支持计划“青年人才”专项(YNWR-QNBJ-2020-154)。

摘　　要：为快速、灵敏、准确检测猫传染性腹膜炎病毒(feline infectious peritonitis virus,FIPV),以FIPV N基因为靶序列设计合成特异性引物及TaqMan探针,建立了一种FIPV实时荧光定量RT-PCR检测方法,并对该方法的特异性、灵敏度和重复性进行了试验。结果表明:该方法灵敏度高,最低检测限为3.4 copies/μL;特异性强,与狂犬病毒、猫疱疹病毒、猫杯状病毒、猫细小病毒、犬流感病毒、犬瘟热病毒等均无交叉反应;重复性好,批内变异系数为0.99%~4.13%,批间变异系数为1.29%~4.64%。以上结果说明,本试验建立的实时荧光定量RT-PCR检测方法特异、敏感、重复性好,适用于FIPV的快速检测。In order to detect feline infectious peritonitis virus(FIPV)quickly,sensitively and accurately,specific primers and TaqMan probes were designed and synthesized based on the FIPV N gene registered in GenBank to establish a real-time fluorescence quantitative RT-PCR assay,followed by the tests for its specificity,sensitivity and repeatability.It was found that,for the established method,high sensitivity and lowest detection limit of 3.4 copies/μL were observed;no cross reaction with rabies virus,feline herpes virus,feline calicivirus,feline parvovirus,canine influenza virus or canine distemper virus was observed due to its strong specificity;and the intra and inter batches of variable coefficients were from 0.99%to 4.13%and from 1.29%to 4.64%,respectively,due to its good repeatability.It was concluded that the established method was appropriate for detecting FIPV rapidly with the help of its good specificity,sensitivity and repeatability.

关 键 词：猫传染性腹膜炎病毒 N基因 TAQMAN探针 荧光定量PCR 检测方法 

分 类 号：S855.3[农业科学—临床兽医学]